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As a
secondary school student you will be testing for the presence of protein in foods.
Instead of the Biuret Reagent, either of the following may be used:
The Biuret test for proteins may also be extended to quantitatively measure
theconcentration of total protein using spectrometric methods.
PROCEDURE
Add 2 cm3 of the liquid food sample* to a clean, dry test tube
Alternatively
Use sodium hydroxide solution and copper sulphate solution instead. Add 1 cm3 of
sodium hydroxide solution (40% or bench solution) and 1% copper (II) sulphate
solution dropwise drop by drop - to the food sample.
OR
Repeat steps 1 and 2 with de-ionized water to prepare a negative control and
with albumin (egg white) to prepare a positive control.
OBSERVATIONS
INTERPRETATION
DISCUSSION QUESTIONS
Q. What is the Principle of the Biuret Test for Proteins?
The Biuret test is based on the ability of Cu (II) ions to form a
violet-coloured chelate
complex with peptide bonds (-CONHgroups) in alkaline conditions.
.
Formation of the Chelate Complex
Lone electron pairs from 4 nitrogen atoms in the peptide bond coordinate a copper (II) ion.
A chelate complex is formed; the complex absorbs light at 540 nm so appears violet. Hence
a color change from blue to violet indicates that proteins are present.
The greater the concentration of peptide bonds, the greater the color intensity. If the
concentration of peptide bonds is low such as when short-chain peptides are present - the
color change is from blue to pink.